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Lipids

, Volume 32, Issue 4, pp 363–367 | Cite as

Nuclear magnetic resonance characterization of 6α-chloro-5β-cholestane-3β,5-diol formed from the reaction of hypochlorous acid with cholesterol

  • Anitra C. Carr
  • Christine C. Winterbourn
  • John W. Blunt
  • Andrew J. Phillips
  • Andrew D. Abell
Article

Abstract

Hyopochlorous acid generated by neutrophil myeloperoxidase has been shown to convert cholesterol into three different chlorohydrin isomers which previously had not been fully characterized. We have reacted hypochlorous acid with cholesterol/1,2-dipalmitoyl phosphatidylcholine liposomes to give these three major products and established that they are 6β-chloro-5α-cholestane-3β,5-diol (chlorohydrin 1), 5α-chloro-6β-cholestane-3β,6-diol (chlorohydrin 2) and 6α-chloro-5β-cholestane-3β,5-diol (chlorohydrin 3). These products were separated by thin-layer chromatography and fully characterized by 1H, 13C, attached proton test, doublequantum correlation spectroscopy, total correlation spectroscopy, heteronuclear multiple bond correlation and heteronuclear multiple quantum coherence nuclear magnetic resonance spectroscopy.

Keywords

DPPC HOCl Hypochlorous Acid Heteronuclear Multiple Bond Correlation Chlorohydrin 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations

APT

attached proton test

DPPC

1,2-dipalmitoyl phosphatidylcholine

DQCOSY

doublequantum correlation spectroscopy

GC-MS

gas chromatography-mass spectrometry

HMBC

heteronuclear multiple bond correlation spectroscopy

HMQC

heteronuclear multiple quantum coherence spectroscopy

NMR

nuclear magnetic resonance

PBS

phosphate-buffered saline

TLC

thin-layer chromatography

TOCSY

total correlation spectroscopy

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References

  1. 1.
    Winterbourn, C.C. (1990) Neutrophil Oxidants: Products and Reactions (Chapter 2), in Oxygen Radicals: Systemic Events and Disease Processes (Das, D.K., and Essman, W.B., eds.) pp. 31–70, Karger, Basel.Google Scholar
  2. 2.
    Prutz, W.A. (1996) Hypochlorous Acid Interactions with Thiols, Nucleotides, DNA, and Other Biological Substrates, Arch. Biochem. Biophys. 332, 110–120.PubMedCrossRefGoogle Scholar
  3. 3.
    Winterbourn, C.C. (1985) Comparative Reactivities of Various Biological Compounds with Myeloperoxidase-Hydrogen Peroxide-Chloride, and Similarity of the Oxidant to Hypochlorite, Biochim. Biophys. Acta, 840, 204–210.PubMedGoogle Scholar
  4. 4.
    Albrich, J.M., McCarthy, C.A., and Hurst, J.K. (1981) Biological Reactivity of Hypochlorous Acid: Implications for Microbicidal Mechanisms of Leukocyte Myeloperoxidase, Proc. Natl. Acad. Sci. USA 78, 210–214.PubMedCrossRefGoogle Scholar
  5. 5.
    Winterbourn, C.C., van den Berg, J.J.M., Roitman, E., and Kuypers, F.A. (1992) Chlorohydrin Formation from Unsaturated Fatty Acids Reacted with Hypochlorous Acid, Arch. Biochem. Biophys. 296, 547–555.PubMedCrossRefGoogle Scholar
  6. 6.
    van den Berg, J.J.M., Winterbourn C.C., and Kuypers, F.A. (1993) Hypochlorous Acid-Mediated Oxidation of Cholesterol and Phospholipid: Analysis of Reaction Products by Gas Chromatography-Mass Spectrometry, J. Lipid Res., 34, 2004–2012.Google Scholar
  7. 7.
    Heinecke, J.W., Li, W., Mueller, D.M., Bohrer, A., and Turk, J. (1994) Cholesterol Chlorohydrin Synthesis by the Myeloperoxidase-Hydrogen Peroxide-Chloride System: Potential Markers for Lipoproteins Oxidatively Damaged by Phagocytes, Biochemistry 33, 10127–10136.PubMedCrossRefGoogle Scholar
  8. 8.
    Carr, A.C., van den Berg, J.J.M., and Winterbourn, C.C. (1996) Chlorination of Cholesterol in Cell Membranes by Hypochlorous Acid, Arch. Biochem. Biophys. 332, 63–69.PubMedCrossRefGoogle Scholar
  9. 9.
    Hazell, L.J., van den Berg, J.J., and Stocker, R. (1994) Oxidation of Low-Density Lipoprotein by Hypochlorite Causes Aggregation That Is Mediated by Modification of Lysine Residues Rather Than Lipid Oxidation, Biochem. J. 302, 297–304.PubMedGoogle Scholar
  10. 10.
    Lindgren, B.O. (1967) Reactions of Sterols with Bleaching Agents: Reactions of Cholesterol and Its Acetate with Aqueous Chlorine Solutions, Svensk Papperstidning 70, 532–536.Google Scholar
  11. 11.
    Lindgren, B.O. (1967) Chlorination of Cholesterol in Aqueous Solution: Isolation of a trans-Diequatorial Chlorohydrin, Acta. Chem. Scand. 21, 1397–1398.Google Scholar
  12. 12.
    Kettle, A.J., and Winterbourn, C.C. (1994) Assays for the Chlorination Activity of Myeloperoxidase, Methods Enzymol. 233, 502–512.PubMedCrossRefGoogle Scholar
  13. 13.
    Hazen, S.L., Hsu, F.F., Duffin, K., and Heinecke, J.W. (1996) Molecular Chlorine Generated by the Myeloperoxidase-Hydrogen Peroxide-Chloride System of Phagocytes Converts Low Density Lipoprotein Cholesterol into a Family of Chlorinated Sterols, J. Biol. Chem. 271, 23080–23088.PubMedCrossRefGoogle Scholar
  14. 14.
    Carr, A.C., Winterbourn, C.C., and van den Berg, J.J.M. (1996) Peroxidase-Mediated Bromination of Unsaturated Fatty Acids to Form Bromohydrins, Arch. Biochem. Biophys. 327, 227–233.PubMedCrossRefGoogle Scholar
  15. 15.
    Blunt, J.M., and Stothers, J.B. (1977) 13C NMR Spectra of Steroids—A Survey and Commentary, Organic Magnetic Resonance 9, 439–464.CrossRefGoogle Scholar
  16. 16.
    Nittala, S.S., Velde, V.V., Frolow, F., and Lavie, D. (1981) Chlorinated Withanolides from Withania somnifera and Acnistus breviflorus, Phytochemistry 20, 2547–2552.CrossRefGoogle Scholar
  17. 17.
    Ali, A., Sahai, M., and Ray, A.B. (1984) Physalolactone C, a New Withanolide from Physalis peruviana, J. Nat. Prod. 47, 648–651.CrossRefGoogle Scholar
  18. 18.
    Smith, L.L., and Johnson, B.H. (1989) Biological Activities of Oxysterols, Free Rad. Biol. Med., 7, 285–332.PubMedCrossRefGoogle Scholar

Copyright information

© AOCS Press 1997

Authors and Affiliations

  • Anitra C. Carr
    • 2
  • Christine C. Winterbourn
    • 2
  • John W. Blunt
    • 1
  • Andrew J. Phillips
    • 1
  • Andrew D. Abell
    • 1
  1. 1.Department of ChemistryUniversity of CanterburyChristchurchNew Zealand
  2. 2.Department of PathologyChristchurch School of MedicineChristchurchNew Zealand

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